High-Performance Liquid Chromatographic Quantification of the Plant Hormone Abscisic Acid at ppb Levels in Plant Samples after a Single Immunoaffinity Column Cleanup.

High-performance liquid chromatography with ultraviolet detection (HPLC-UV) is a common analysis technique due to its high versatility and simple operation. In the present study, HPLC-UV detection was integrated with immunoaffinity cleanup (IAC) of the sample extracts. The matrix effect was greatly reduced, and the limit of detection was as low as 1 ng/g of free abscisic acid (ABA) in fresh plant tissues. A monoclonal antibody 3F1 (mAb 3F1) was developed to specifically recognize free ABA but not ABA analogues. The mAb 3F1-immobilized immunoaffinity column exhibited a capacity of 850 ng/mL and an elution efficiency of 88.8-105% for standards. The extraction recoveries of the column for ABA ranged from 80.4 to 108.9%. ABA content was detected in various plant samples with IAC-HPLC-UV. The results were verified with ultraperformance liquid chromatography-electrospray tandem mass spectrometry. IAC-HPLC-UV can be a sensitive and cost-efficient method for plant hormone analysis.

Hormonal profiles in dormant turions of 22 aquatic plant species: do they reflect functional or taxonomic traits?

BACKGROUND AND AIMS: Turions are vegetative, dormant overwintering organs formed in aquatic plants in response to unfavourable ecological conditions. Contents of cytokinin (CK), auxin metabolites and abscisic acid (ABA) as main growth and development regulators were compared in innately dormant autumnal turions of 22 aquatic plant species of different functional ecological or taxonomic groups with those in non-dormant winter apices in three aquatic species and with those in spring turions of four species after their overwintering. METHODS: The hormones were analysed in miniature turion samples using ultraperformance liquid chromatography coupled with triple quadrupole mass spectrometry. KEY RESULTS: In innately dormant turions, the total contents of each of the four main CK types, biologically active forms and total CKs differed by two to three orders of magnitude across 22 species; the proportion of active CK forms was 0.18-67 %. Similarly, the content of four auxin forms was extremely variable and the IAA proportion as the active form was 0.014-99 %. The ABA content varied from almost zero to 54 µmol kg-1 dry weight and after overwintering it usually significantly decreased. Of all functional traits studied, hormone profiles depended most on the place of turion sprouting (surface vs bottom) and we suggest that this trait is crucial for turion ecophysiology. CONCLUSIONS: The key role of ABA in regulating turion dormancy was confirmed. However, the highly variable pattern of the ABA content in innately dormant and in overwintered turions indicates that the hormonal mechanism regulating the innate dormancy and its breaking in turions is not uniform within aquatic plants.

A rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry method for determination of phytohormones in the medicinal plant saffron.

Saffron (Crocus sativus L.) is a valuable Chinese herb with high medicinal value. Saffron pistils are used as medicine, so increasing the number of flowers can increase the yield. Plant hormones have essential roles in the growth and development of saffron, as well as the response to biotic and abiotic stresses (especially in floral initiation), which may directly affect the number of flowers. Quantitative analysis of plant hormones provides a basis for more efficient research on their synthesis, transportation, metabolism, and action. However, starch (which interferes with extraction) is present in high levels, and hormone levels are extremely low, in saffron corms, thereby hampering accurate determination of plant-hormone levels in saffron. Herein, we screened an efficient and convenient pre-treatment method for plant materials containing abundant amounts of starch. Also, we proposed an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the quantification of abscisic acid (ABA) and auxin (IAA). Then, the method was applied for the detection of hormone-content differences between flowering and non-flowering top buds, as well as between lateral and top buds. Our method showed high sensitivity, reproducibility, and reliability. Specifically, good linearity in the range 2-100 ng ml-1 was achieved in the determination of ABA and IAA, and the correlation coefficient (R2) was >0.9982. The relative standard deviation was 2.956-14.51% (intraday) and 9.57-18.99% (interday), and the recovery range was 89.04-101.1% (n = 9). The matrix effect was 80.38-90.50% (n = 3). The method was thoroughly assessed employing various "green" chemistry evaluation tools: Blue Applicability Grade Index (BAGI), Complementary Green Analytical Procedure Index (Complex GAPI) and Red Green Blue 12 Algorithm (RGB12). These tools revealed the good greenness, analytical performance, applicability, and overall sustainability alignment of our method. Quantitative results showed that, compared with saffron with a flowering phenotype cultivated at 25 °C, the contents of IAA and ABA in the terminal buds of saffron cultivated at 16 °C decreased significantly. When cultivated at 25 °C, the IAA and ABA contents in the terminal buds of saffron were 1.54- and 4.84-times higher than those in the lateral buds, respectively. A simple, rapid, and accurate UPLC-MS/MS method was established to determine IAA and ABA contents. Using this method, a connection between the contents of IAA and ABA and the flowering phenotype was observed in the quantification results. Our data lay a foundation for studying the flowering mechanism of saffron.

NRT1.2 overexpression enhances the synergistic interplay between ABA-generating bacteria and biochars in reducing heavy metal accumulation in pak choi.

The agricultural sector faces severe challenges owing to heavy metal (HM) contamination of farmlands, requiring urgent preventive measures. To address this, we investigated the impact of the synergistic application of Azospirillum brasilense, a growth-promoting rhizobacterium producing abscisic acid (ABA), and biochar to minimize HM accumulation in pak choi, using three distinct expression levels of the ABA transporter NRT1.2 in pak choi and three different types of contaminated soils as experimental materials. The results revealed that pak choi with low, medium, and high NRT1.2 expression intensity, when subjected to bacterial strain-biochar treatment, exhibited an increasing trend in ABA content compared to the control. Correspondingly, the aboveground HM content decreased by 1-49 %, 22-52 %, and 15-96 %, whereas the fresh weight increased by 12-38 %, 88-126 %, and 152-340 %, respectively, showing a significant correlation with NRT1.2 expression. Pearson correlation analysis demonstrated that NRT1.2 expression intensity was inversely associated with the combined treatment's reduction in HM accumulation and positively correlated with the promotional effect. Simultaneously, soil discrepancies significantly affected the combined treatment, which was likely associated with variations in the active forms of HM in each soil. Consequently, when employing ABA-producing bacteria for mitigating crop HM accumulation, selecting plants with higher relative NRT1.2 expression intensity, combined with biochar, is recommended.

An arbuscular mycorrhizal fungus differentially regulates root traits and cadmium uptake in two maize varieties.

Arbuscular mycorrhizal fungi (AMF) are symbiotic fungi that colonize plant roots, and they are more common in Cd-polluted habitats. However, there is limited understanding of the response of root traits and cadmium (Cd) uptake to AMF in different crop varieties. Two maize varieties, Panyu 3 and Ludan 8, with high and low Cd uptake capacities, respectively, were cultivated as host plants in a pot experiment with Cd-polluted soil (17.1 mg/kg Cd). The effects of AMF on the growth, mineral nutrient concentration, root traits, phytohormone concentrations and Cd uptake of the two maize varieties and their comprehensive response to AMF fungal inoculation were investigated. AMF improved growth, mineral nutrient levels and root morphology and increased lignin and phytohormone concentrations in roots and Cd uptake in the two maize varieties. However, the two maize varieties, Panyu 3 and Ludan 8, had different responses to AMF, and their comprehensive response indices were 753.6% and 389.4%, respectively. The root biomass, branch number, abscisic acid concentrations, lignin concentrations and Cd uptake of maize Panyu 3 increased by 151.1%, 28.6%, 139.7%, 99.5% and 84.7%, respectively. The root biomass, average diameter, auxin concentration, lignin concentration and Cd uptake of maize Ludan 8 increased by 168.7%, 31.8%, 31.4%, 41.7% and 136.7%, respectively. Moreover, Cd uptake in roots presented very significant positive correlations with the average root diameter and abscisic acid concentration. A structural equation model indicated that the root abscisic acid concentration and root surface area had positive effects on Cd uptake by the Panyu 3 maize roots; the root abscisic acid concentration and root tip number had positive effects on Cd uptake by the Ludan 8 maize roots. Thus, AMF differentially regulated Cd uptake in the two maize varieties, and the regulatory effect was closely related to root traits and phytohormone concentrations.

ABA-metabolizing bacteria and rhamnolipids as valuable allies for enhancing phytoremediation efficiency in heavy metal-contaminated soils.

Microbial-assisted phytoremediation has great potential to improve the efficiency of phytoremediation in heavy metal (HM)-contaminated soils. In this study, the synergistic effects of rhamnolipids and the abscisic acid (ABA)-metabolizing bacterium Rhodococcus qingshengii on the phytoremediation efficiency of Indian mustard (Brassica juncea) in HM-contaminated soils were investigated. The Cd, Zn, and Pb contents in plants treated with a combination of rhamnolipids and R. qingshengii were 48.4-77.1 %, 14.6-40.4 %, and 16.1-20.0 % higher, respectively, than in those treated with R. qingshengii alone, and 42.8-59.2 %, 13.1-48.2 %, and 7.3-67.5 % higher, respectively, than in those treated with rhamnolipids alone. In addition, the bioconcentration factors of each metal were improved, and the biomass further increased by 36.6-65.7 % compared to that of single treatments. Pearson's correlation analysis showed that rhamnolipids and R. qingshengii enhanced the accumulation of HMs in B. juncea by activating the available forms of HMs in the soil and regulating the ABA and indole-3-acetic acid in plants, respectively. The structural equation model indicated that R. qingshengii had a larger path coefficient than rhamnolipids in terms of HM content and plant biomass, suggesting that R. qingshengii may have a greater contribution to promoting the extraction of HMs from the soil under synergistic conditions. In conclusion, the combination of rhamnolipids and R. qingshengii has great potential to enhance the phytoremediation efficiency of hyperaccumulating plants in HM-contaminated soils.

A Novel, Simple Rapid Reverse-Phase HPLC-DAD Analysis, for the Simultaneous Determination of Phenolic Compounds and Abscisic Acid Commonly Found in Foodstuff and Beverages.

A novel, simple, rapid, 7-minute HPLC-DAD method for the determination of 10 phenolic compounds and abscisic acid commonly found in teas, wines, fruit and honey was successfully developed and validated according to the International Council of Harmonization (ICH) guidelines. This reverse-phase (RP) HPLC-DAD method boasts rapid separation and excellent resolution while introducing green chemistry techniques. The Agilent 1200 series diode array detector SL coupled with a reverse-phase Advanced Materials Technology Halo C18 column (100 × 3.0 mm i.d., 2.7 µm) contributed to the rapid analyses. This, together with a 0.1% formic acid in water (v/v) and methanol mobile phase, a flow rate of 0.8 mL/min and the utilization of a meticulous gradient elution resulted in a validated method for the determination of 10 phenolic compounds and abscisic acid commonly found in various foodstuffs. The resulting method proved to be rapid, accurate, precise and linear with sensitive detection limits from 0.025 µg/mL to 0.500 µg/mL and percentage recoveries of 98.07%-101.94%. Phenolic compounds have been acknowledged throughout literature for their therapeutic properties, interalia, antioxidant, anti-inflammatory and antiaging due to free radical scavenging potentials. However, resulting analysis, can be frequently complicated and long and very often discounts green chemistry techniques. The developed and validated method successfully and rapidly analyses, gallic acid, caffeic acid, trans-p-coumaric acid, rutin, myricetin, abscisic acid, trans-cinnamic acid, quercetin, luteolin, kaempferol and chrysin with excellent resolution and precision.

Uncovering the effects of kaolin on balancing berry phytohormones and quality attributes of Vitis vinifera grown in warm-temperate climate regions.

BACKGROUND: The application of kaolin particle film is considered a short-term strategy against several environmental stresses in areas with a Mediterranean-like climate. However, it is known that temperature fluctuations and water availability over the season can jeopardize kaolin efficiency in many Mediterranean crops. Hence, this study aims to evaluate the effects of kaolin foliar application on berry phytohormones, antioxidant defence, and oenological parameters at veraison and harvest stages of Touriga-Franca (TF) and Touriga-Nacional (TN) grapevines in two growing seasons (2017 and 2018). The 2017 growing season was considered the driest (-147.1 dryness index) and the warmest (2705 °C growing degree days) of the study. RESULTS: In 2017, TF kaolin-treated berries showed lower salicylic acid (-26.6% compared with unsprayed vines) and abscisic acid (ABA) (-10.5%) accumulation at veraison, whereas salicylic acid increased up to 28.8% at harvest. In a less hot season, TN and TF kaolin-treated grapevines showed a twofold in ABA content and a threefold increase in the indole-3-acetic acid content at veraison and lower ABA levels (83.8%) compared with unsprayed vines at harvest. Treated berries showed a decreased sugar content, without compromising malic and tartaric acid levels, and reactive oxygen species accumulation throughout berry ripening. CONCLUSION: The results suggest kaolin exerts a delaying effect in triggering ripening-related processes under severe summer stress conditions. Treated berries responded with improved antioxidant defence and phytohormone balance, showing significant interactions between kaolin treatment, variety, and developmental stage in both assessed years. © 2021 Society of Chemical Industry.

Phytohormone profiles in non-transformed and AtCKX transgenic centaury (Centaurium erythraea Rafn) shoots and roots in response to salinity stress in vitro.

Plant hormones regulate numerous developmental and physiological processes. Abiotic stresses considerably affect production and distribution of phytohormones as the stress signal triggers. The homeostasis of plant hormones is controlled by their de novo synthesis and catabolism. The aim of this work was to analyse the contents of total and individual groups of endogenous cytokinins (CKs) as well as indole-3-acetic acid (IAA) in AtCKX overexpressing centaury plants grown in vitro on graded NaCl concentrations (0, 50, 100, 150, 200 mM). The levels of endogenous stress hormones including abscisic acid (ABA), salicylic acid (SA) and jasmonic acid (JA) were also detected. The elevated contents of total CKs were found in all analysed centaury shoots. Furthermore, increased amounts of all five CK groups, as well as enhanced total CKs were revealed on graded NaCl concentrations in non-transformed and AtCKX roots. All analysed AtCKX centaury lines exhibited decreased amounts of endogenous IAA in shoots and roots. Consequently, the IAA/bioactive CK forms ratios showed a significant variation in the shoots and roots of all AtCKX lines. In shoots and roots of both non-transformed and AtCKX transgenic centaury plants, salinity was associated with an increase of ABA and JA and a decrease of SA content.

Complex Analysis of Antioxidant Activity, Abscisic Acid Level, and Accumulation of Osmotica in Apple and Cherry In Vitro Cultures under Osmotic Stress.

Plant response to osmotic stress is a complex issue and includes a wide range of physiological and biochemical processes. Extensive studies of known cultivars and their reaction to drought or salinity stress are very important for future breeding of new and tolerant cultivars. Our study focused on the antioxidant activity, accumulations of osmotica, and the content of abscisic acid in apple (cv. "Malinové holovouské", "Fragrance", "Rubinstep", "Idared", "Car Alexander") and cherry (cv. "Regina", "Napoleonova", "Kastánka", "Sunburst", "P-HL-C") cultivated in vitro on media containing different levels of polyethylene glycol PEG-6000. Our results indicated that the studied genotypes responded differently to osmotic stress manifested as reduction in the leaf relative water content (RWC) and increment in the activities of antioxidant enzymes, proline, sugars, and abscisic acid content. Overall, cherry cultivars showed a smaller decrease in percentage RWC and enzymatic activities, but enhanced proline content compared to the apple plants cultivars. Cultivars "Rubinstep", "Napoleonova", and "Kastánka" exhibited higher antioxidant capacity and accumulation of osmoprotectants like proline and sorbitol that can be associated with the drought-tolerance system.

Flooding or drought which one is more offensive on pepper physiology and growth?

Both extreme usage of water in agriculture i.e., drought and flooding affect physiological and growth aspects of the plant as well as gene expression undertaken in water absorption. These affect depend on the stress duration i.e., shock or gradual stress exposer. The factorial experiment based on CRD with 10 replicates was conducted to investigate the physiological and water relation as well as aquaporin expression in (Capsicum annuum L.). Drought stress was applied gradually from - 2, - 3, - 4 to - 5 MPa during 8 days but in shock stress - 5 MPa applied at one time. The gradual flooding stress adjusted with changing the aeration duration from 15 to 0 min gradually every 2 days and for the shock- flooding, peppers keep in a nutrient solution without aeration in a sealed container. Results showed that both extreme water stress had a deleterious effect on the growth and physiological parameter of pepper for a longer duration. Antioxidant, proline, fluorescence chlorophyll stimulate in the gradual period except for ABA content, which is higher in shock stress. PIP1expression showed a reverse effect in leaf and root at flooding i.e., PIP1expression raised in root while it was reduced in leaf at shock-flooding. The highest PIP1expression was observed in gradual-drought of root and gradual duration of drought and flooding stress in leaf. In the physiological aspect of plant response to stress in pepper, results showed an enhanced in proline and phenol content to help osmotic adjustment and keep water status in moderate condition. Conclusively, shocked stress first, motivated these defense systems, and then in the next step, the other adaptive mechanism like gene expression activated to help pepper face stress. On the other hand, shock stress showed down-regulation, but when the stress lasted for a longer time results in up-regulation.

In vivo FRET-FLIM reveals ER-specific increases in the ABA level upon environmental stresses.

Plant hormone abscisic acid (ABA) is essential for regulating plant growth and various stress responses. ABA-mediated signaling depends on local ABA levels rather than the overall cellular ABA concentration. While cellular concentration of ABA can be detected using Förster resonance energy transfer (FRET)-based ABA probes, direct imaging of subcellular ABA levels remains unsolved. Here, we modified the previously reported ABAleon2.1 and generated a new ABA sensor, named ABAleon2.1_Tao3. Via transient expression in tobacco (Nicotiana tabacum) protoplasts, we targeted ABAleon2.1_Tao3s to the endoplasmic reticulum (ER) membrane with the ABA sensing unit facing the cytosol and the ER, respectively, through a nanobody-epitope-mediated protein interaction. Combining FRET with fluorescence lifetime imaging microscopy, ABA-triggered-specific increases in the fluorescence lifetime of the donor mTurquoise in the ABAleon2.1_Tao3 were detected in both transient assays and stably transformed Arabidopsis plants. In tobacco protoplasts, ER membrane-targeted ABAleon2.1_Tao3s showed a generally higher basal level of ABA in the ER than that in the cytosol and ER-specific alterations in the level of ABA upon environmental cues. In ABAleon2.1_Tao3-transformed Arabidopsis roots, mannitol triggered increases in cytosolic ABA in the division zone and increases in ER ABA in the elongation and maturation zone within 1 h after treatment, both of which were abolished in the bg1-2 mutant, suggesting the requirement for BG1 in osmotic stress-triggered early ABA induction in Arabidopsis roots. These data demonstrate that ABAleon2.1_Tao3s can be used to monitor ABA levels in the cytosol and the ER, providing key information on stress-induced changes in the level of ABA in different subcellular compartments.

Mitigation of the effect of drought on growth and yield of pomegranates by foliar spraying of different sizes of selenium nanoparticles.

BACKGROUND: Drought is a very important environmental stressor, which has negative effects on the growth of trees, decreasing their yield. The role of different-sized selenium nanoparticles (Se-NPs) in the mitigation of environmental stresses such as drought in crops has not yet been investigated. RESULTS: Trees treated with Se-NPs displayed higher levels of photosynthetic pigments, a better nutrient status, better physical parameters (especially fruit cracking) and chemical parameters, a higher phenolic content, and higher concentrations of osmolytes, antioxidant enzymes, and abscisic acid than untreated trees under drought stress. Foliar spraying of 10 and 50 nm Se-NPs alleviated many of the deleterious effects of drought in pomegranate leaves and fruits and this was achieved by reducing stress-induced lipid peroxidation and H2 O2 content by enhancing the activity of antioxidant enzymes. Furthermore, the 10 nm Se-NPs treatment produced more noticeable effects than the treatment with 50 nm Se-NPs. CONCLUSION: Results confirm the positive effects of nanoparticle spraying, especially the role of 10 nm Se-NPs in the management of negative effects of drought not only for pomegranates but potentially also for other crops. © 2021 Society of Chemical Industry.

Sensors for the quantification, localization and analysis of the dynamics of plant hormones.

Plant hormones play important roles in plant growth and development and physiology, and in acclimation to environmental changes. The hormone signaling networks are highly complex and interconnected. It is thus important to not only know where the hormones are produced, how they are transported and how and where they are perceived, but also to monitor their distribution quantitatively, ideally in a non-invasive manner. Here we summarize the diverse set of tools available for quantifying and visualizing hormone distribution and dynamics. We provide an overview over the tools that are currently available, including transcriptional reporters, degradation sensors, and luciferase and fluorescent sensors, and compare the tools and their suitability for different purposes.

Changes in the distribution of endogenous hormones in Phyllostachys edulis 'Pachyloen' during bamboo shooting.

In this study, we investigated the changes in the distribution and regulation of endogenous hormones in Phyllostachys edulis 'Pachyloen' during bamboo shooting. Enzyme-linked immunosorbent assay was used to measure the mass fractions of indole-3-acetic acid (IAA), gibberellic acid (GA), zeatin riboside (ZR), and abscisic acid (ABA) in rhizomes, shoots, and maternal bamboo organs during shoot sprouting, shoot growth, and new-bamboo formation. Measurements were compared among bamboo parts and developmental periods. The overall mass fractions of IAA and ABA were significantly higher than those of ZR and GA, driven by differences among bamboo parts and developmental periods. The abundance of each endogenous hormone varied among bamboo parts and developmental periods. During bamboo shooting, ABA had the highest mass fraction in all bamboo parts sampled, followed by IAA, GA, and ZR. Among bamboo parts, rhizomes had more IAA, ZR, and GA than the other parts, but significantly less ABA. Winter shoots had higher ZR: IAA and GA: IAA ratios than rhizomes and maternal bamboo organs. During shoot growth, ABA was the most abundant hormone in rhizomes and maternal bamboo organs, followed by IAA, ZR, and GA. In contrast, IAA was the most abundant hormone in spring shoots, followed by ABA, ZR, and GA. Maternal bamboo organs had a significantly higher ZR: GA ratio, and significantly lower IAA: ABA, ZR: ABA, and GA: ABA ratios than rhizomes. Spring shoots had significantly higher IAA: ABA, ZR: ABA, and GA: ABA ratios than rhizomes and maternal bamboo organs; significantly higher ZR mass fractions, and ZR: GA and ZR: IAA ratios and significantly lower ABA mass fractions than rhizomes; and significantly higher GA: IAA ratio than maternal bamboo organs. During new-bamboo formation, ABA was the most abundant hormone in rhizomes, winter shoots, and maternal bamboo organs, followed by IAA, ZR, and GA. Maternal bamboo organs had significantly lower IAA mass fractions and significantly higher ABA mass fractions than rhizomes and new bamboo tissue. IAA and ABA abundances exhibited an inverse relationship in rhizomes and maternal bamboo organs. GA: ABA and GA: IAA ratios decreased gradually and other hormone ratios exhibited parabolic trends over the bamboo-shooting period, with the highest ratios observed in new bamboo tissues. Overall, the coordination or antagonism among endogenous hormones plays a key regulatory role in bamboo shoot growth. The formation of thick walls in P. edulis 'Pachyloen', one of its major traits, may be partially attributed to the relatively high IAA and ZR and low GA mass fractions.

UPLC-QTof-MSE Metabolomics Reveals Changes in Leaf Primary and Secondary Metabolism of Hop (Humulus lupulus L.) Plants under Drought Stress.

The hop (Humulus lupulus L.) is an important specialty crop used in beer production. Untargeted UPLC-QTof-MSE metabolomics was used to determine metabolite changes in the leaves of hop plants under varying degrees of drought stress. Principal component analysis revealed that drought treatments produced qualitatively distinct changes in the overall chemical composition of three out of four genotypes tested (i.e., Cascade, Sultana, and a wild var. neomexicanus accession but not Aurora), although differences among treatments were smaller than differences among genotypes. A total of 14 compounds consistently increased or decreased in response to drought stress, and this effect was generally progressive as the severity of drought increased. A total of 10 of these marker compounds were tentatively identified as follows: five glycerolipids, glutaric acid, pheophorbide A, abscisic acid, roseoside, and dihydromyricetin. Some of the observed metabolite changes likely occur across all plants under drought conditions, while others may be specific to hops or to the type of drought treatments performed.

Multiomic Analysis Elucidates the Reasons Underlying the Differential Metabolite Accumulation in Citrus Mature Leaves and Fruit Juice Sacs.

Fruit and leaf possess distinctly different metabolites. Here, metabolites and transcriptome were compared between mature leaves (ML) and juice sacs (JS) of Citrus grandis "Hirado Buntan" to investigate the possible reasons. Results indicated that the remarkable difference in starch, total flavonoids and carotenoids, l-ascorbate, and jasmonic acid between ML and JS was tightly related to the expression levels of their biosynthesis-related genes, while the significant difference in abscisic acid and citrate was mainly related to the gene expression level(s) of 9-cis-epoxycarotenoid dioxygenase and proton pump genes, respectively. In addition, ATP citrate lyase probably plays a key role in the levels of flavonoids between ML and JS via providing different levels of acetyl-CoA. Taken together, these results identified some key candidate genes responsible for the content of a given metabolite and will contribute to research in regulating such metabolite content in citrus fruits.

Transcription profiles reveal sugar and hormone signaling pathways mediating tree branch architecture in apple (Malus domestica Borkh.) grafted on different rootstocks.

Apple trees grafted on different rootstock types, including vigorous rootstock (VR), dwarfing interstock (DIR), and dwarfing self-rootstock (DSR), are widely planted in production, but the molecular determinants of tree branch architecture growth regulation induced by rootstocks are still not well known. In this study, the branch growth phenotypes of three combinations of 'Fuji' apple trees grafted on different rootstocks (VR: Malus baccata; DIR: Malus baccata/T337; DSR: T337) were investigated. The VR trees presented the biggest branch architecture. The results showed that the sugar content, sugar metabolism-related enzyme activities, and hormone content all presented obvious differences in the tender leaves and buds of apple trees grafted on these rootstocks. Transcriptomic profiles of the tender leaves adjacent to the top buds allowed us to identify genes that were potentially involved in signaling pathways that mediate the regulatory mechanisms underlying growth differences. In total, 3610 differentially expressed genes (DEGs) were identified through pairwise comparisons. The screened data suggested that sugar metabolism-related genes and complex hormone regulatory networks involved the auxin (IAA), cytokinin (CK), abscisic acid (ABA) and gibberellic acid (GA) pathways, as well as several transcription factors, participated in the complicated growth induction process. Overall, this study provides a framework for analysis of the molecular mechanisms underlying differential tree branch growth of apple trees grafted on different rootstocks.

Genome-wide characterization of the abscisic acid-, stress- and ripening-induced (ASR) gene family in wheat (Triticum aestivum L.).

BACKGROUND: Abscisic acid-, stress-, and ripening-induced (ASR) genes are a class of plant specific transcription factors (TFs), which play important roles in plant development, growth and abiotic stress responses. The wheat ASRs have not been described in genome-wide yet. METHODS: We predicted the transmembrane regions and subcellular localization using the TMHMM server, and Plant-mPLoc server and CELLO v2.5, respectively. Then the phylogeny tree was built by MEGA7. The exon-intron structures, conserved motifs and TFs binding sites were analyzed by GSDS, MEME program and PlantRegMap, respectively. RESULTS: In wheat, 33ASR genes were identified through a genome-wide survey and classified into six groups. Phylogenetic analyses revealed that the TaASR proteins in the same group tightly clustered together, compared with those from other species. Duplication analysis indicated that the TaASR gene family has expanded mainly through tandem and segmental duplication events. Similar gene structures and conserved protein motifs of TaASRs in wheat were identified in the same groups. ASR genes contained various TF binding cites associated with the stress responses in the promoter region. Gene expression was generally associated with the expected group-specific expression pattern in five tissues, including grain, leaf, root, spike and stem, indicating the broad conservation of ASR genes function during wheat evolution. The qRT-PCR analysis revealed that several ASRs were up-regulated in response to NaCl and PEG stress. CONCLUSION: We identified ASR genes in wheat and found that gene duplication events are the main driving force for ASR gene evolution in wheat. The expression of wheat ASR genes was modulated in responses to multiple abiotic stresses, including drought/osmotic and salt stress. The results provided important information for further identifications of the functions of wheat ASR genes and candidate genes for high abiotic stress tolerant wheat breeding.

Life-Course Monitoring of Endogenous Phytohormone Levels under Field Conditions Reveals Diversity of Physiological States among Barley Accessions.

Agronomically important traits often develop during the later stages of crop growth as consequences of various plant-environment interactions. Therefore, the temporal physiological states that change and accumulate during the crop's life course can significantly affect the eventual phenotypic differences in agronomic traits among crop varieties. Thus, to improve productivity, it is important to elucidate the associations between temporal physiological responses during the growth of different crop varieties and their agronomic traits. However, data representing the dynamics and diversity of physiological states in plants grown under field conditions are sparse. In this study, we quantified the endogenous levels of five phytohormones - auxin, cytokinins (CKs), ABA, jasmonate and salicylic acid - in the leaves of eight diverse barley (Hordeum vulgare) accessions grown under field conditions sampled weekly over their life course to assess the ongoing fluctuations in hormone levels in the different accessions under field growth conditions. Notably, we observed enormous changes over time in the development-related plant hormones, such as auxin and CKs. Using 3' RNA-seq-based transcriptome data from the same samples, we investigated the expression of barley genes orthologous to known hormone-related genes of Arabidopsis throughout the life course. These data illustrated the dynamics and diversity of the physiological states of these field-grown barley accessions. Together, our findings provide new insights into plant-environment interactions, highlighting that there is cultivar diversity in physiological responses during growth under field conditions.